Method of storing a vaccine containing an aluminum adjuvant

ABSTRACT

The invention relates to a method for loading and storing a vaccine composition, containing the antigen adsorbed on the aluminum adjuvant which (a) comprises (i) loading the composition into a container; and (ii) closing the container with a device in particular acting as a stopper, the surface of the device getting into contact with the composition being coated with a fluoropolymer such as Teflon™; and/or (b) loading the composition into a container wherein the inner surface of which is coated with polymerized silicone. The use of fluoropolymer or polymerized silicone optimizes the adsorbed antigen stability upon storage. In a particular embodiment, the antigen is the hepatitis B surface antigen and the aluminum adjuvant is aluminum oxy hydroxide.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. provisional application61/454,248, filed Mar. 18, 2011, and French patent application no. 1058464, filed Oct. 18, 2011. The entire contents of both of theseapplications are incorporated by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a method for reducing and/ordecelerating the desorption of an antigen that has been adsorbed on analuminum adjuvant as well as to the product thereof—namely thecombination of a vaccine composition comprising the antigen adsorbed onthe aluminum adjuvant and a container for the composition, saidcontainer having particular characteristics.

2. Summary of the Related Art

A large number of antigens are able to adsorb on an aluminum adjuvant,in particular at neutral pH or at a pH close to neutrality, which is thepH naturally required for compositions that have to be administered tomammals, including humans.

Provided that the amount of adjuvant is such that the antigen canactually adsorb on the adjuvant in an optimum amount when the twocompounds are mixed together, the maximum degree of adsorption is veryfrequently achieved. However, over time, depending on the environmentalconditions, the percentage of adsorbed antigen (adsorption rate) maydecrease, and this desorption may constitute an instability factor.

Known environmental conditions that can affect the percentage ofadsorbed antigen (adsorption rate) include, for example, variations inpH (even slight variations), and the addition of one or more mediumcomponent(s) or one or more additional antigen(s) that may compete withthe first antigen for the adsorption sites on the adjuvant.

Conventionally, a ready-to-use multi-dose vaccine composition is loadedinto vials, e.g., glass vials closed with a plastic stopper. Similarly,a single dose of a vaccine composition may be loaded in a mono-dose vialor a ready-for-injection syringe consisting, in a standard manner, of areservoir containing the vaccine, a plunger that closes the reservoir atits distal end, and a device for administration, such as a needleattached at its proximal end. According to an alternative standardfilling mode, the vaccine dose may also be loaded in a needlelesssyringe to which the practitioner adds a separately packaged needle atthe time of the injection. The reservoir of the syringe is generallymade of either glass or plastic and the plunger or the stopper is simplymade of plastic, such as a chlorobutyl or bromobutyl polymer, withoutparticular lamination. Standard glass or plastic syringes are sold, forexample, by Becton-Dickinson; Gerresheimer A G, Schott A G, Germany;Nuova Ompi srl, Italy; and West Pharma/Daykio. In order to facilitatesliding, the plunger or stopper may have been immersed in asilicone-in-water emulsion so that a silicone film is formed at itssurface. Standard plungers/stoppers are sold by Helvoet, Stelmi and WestPharma, for example; some of them already being sold coated with asilicone film (ref. B2 from West Pharma).

Whatever the container used for storage and the device used for closingthe container, administration at the time of injection consists of usinga syringe and sliding the plunger or the stopper/plunger combination sothat the vaccine is delivered.

SUMMARY OF THE INVENTION

We have now found that the material of the container itself as well asthat of the device for closing the container can affect the adsorptionrate.

We observed that a vaccine composition containing the hepatitis Bsurface antigen (HBsAg) adjuvanted with an aluminum adjuvant and loadedas a single dose in standard syringe closed with a standard stopperunderwent different adsorption rates when stored under identicalconditions and for the same period of time (a few days to severalmonths) depending on whether the syringe was stored in the vertical orhorizontal position. The same phenomenon has also been seen with vials.The practical consequence of the different storage positions was thatthe vaccine contained in the reservoirs stored vertically was not incontact with the stopper, whereas there was contact between thecomposition and the stopper in the horizontally stored reservoirs.

After a certain period of time, the level of adsorption of HBsAg wasmeasured, and we observed that the level of adsorption was much lowerfor the HBsAg contained in the horizontally-stored reservoirs. Thisindicated to us that the material of the standard stopper (chlorobutylor bromobutyl polymer) was responsible for the adsorption decrease.

The solution to this problem is either to prevent contact between theadjuvanted vaccine composition and a stopper made of a material thatcontributes to decreased adsorption or to use a stopper made of amaterial that does not contribute to decreased adsorption. In oneembodiment, the invention comprises a method and apparatus using astopper coated with a film of a fluoropolymer, such as Teflon™ or aTeflon™—like substance (such as Omniflex™ from Helvoet Pharma orFluorotec™ from West Pharma), which decrease or eliminate desorptioncaused by the stopper.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the invention comprises a first method for reducingand/or slowing down the desorption of an antigen initially adsorbed onan aluminum adjuvant during storage, the method comprising (i) loading acontainer with a vaccine composition containing the antigen initiallyadsorbed on the aluminum adjuvant; and (ii) closing the container with adevice acting as a stopper, the surface of the device contactable withthe composition being coated with a fluoropolymer.

In other words, the invention relates to a first method for filling andstoring a composition containing the antigen adsorbed on the aluminumadjuvant which comprises (i) filling a container with the vaccinecomposition; and (ii) closing the container with a device acting as astopper, the surface of the device contactable with the compositionbeing coated with a fluoropolymer.

In a similar manner, the invention also relates to the use of a deviceacting as a stopper for closing a container containing a vaccinecomposition comprising the antigen adsorbed on an aluminum adjuvant, thesurface of the device contactable with the composition being coated witha fluoropolymer.

The container may be, for example, a vial or the reservoir of a syringe.This also applies to all the other aspects of the invention describedhereinafter.

The vaccine composition may be liquid or solid, e.g. lyophilized. Alyophilized composition may have the appearance of a powder. At the timeof injection to a patient, the lyophilized composition is reconstitutedwith an appropriate pharmaceutical solution. This also applies to allthe other aspects of the invention described hereinafter.

According to conventional practice in the art and for the purposesherein, it is understood that “antigen adsorbed” or “initially adsorbed”is not intended to mean that 100% of the antigen amount is actuallyadsorbed. These terms simply mean that a substantial amount of antigenis adsorbed. This also applies to all the other aspects of the inventiondescribed hereinafter.

As mentioned above, the device may be a plastic device made out of, forexample, a chlorobutyl or bromobutyl polymer. This standard device istreated with a fluoropolymer; in particular, it may be submitted to alaminar flow treatment with a fluoropolymer, this laminar flow treatmentbeing carried out on the entire device or, at the very least, on thesurface of the device contactable with the composition. The laminar flowtreatment makes it possible to deposit a very thin layer (e.g., film) ofthe fluoropolymer. As will be appreciated, the area of the coatedsurface may exceed the surface contactable with the composition. Indeed,in one embodiment, the whole surface of the device is coated with thefluoropolymer.

For use in the present invention, the fluoropolymer may be, for example,polytetrafluoroethylene (PTFE), polytetrafluoropropylene (PTFP),fluorinated ethylene propylene (FEP, a copolymer of hexafluoropropyleneand tetrafluoroethylene), polychlorotrifluoroethylene (PCTFE),perfluoroalkoxy co-polymer (PFA), poly(ethylene-co-tetrafluoroethylene)(ETFE), poly(ethylenechlorotrifluoroethylene) (ECTFE), polyvinylfluoride (PVF) or polyvinylidene fluoride (PVPF).

The method/use disclosed herein makes it possible to reduce thedesorption speed of the antigen adsorbed on the aluminum adjuvant and/orthe desorption percentage (or desorption rate) after a defined storagetime at a given temperature. The desorption rate may be expressed asfollows: (amount of non-adsorbed antigen)/(total antigen amount presentin the composition). Typically, the desorption rate can be assessed bycentrifuging the vaccine composition (samples at T (time)=0 and at theend of the experiment); recovering the supernatants which contain thedesorbed antigen; and then quantifying the desorbed fraction by assayingthe antigen in the supernatants and in the whole vaccine using asuitable method chosen according to the nature of the antigen. Thedesorption percentage (or desorption rate) can vary from one antigen toanother according to the strength/weakness of the antigen-adjuvantinteraction. Nevertheless, it is considered that the desorptionpercentage (or desorption rate) can be reduced by 10 to 15 or 20%compared with a standard loading method using standard stoppers—saidreduction being measured 1 or 2 months after the date of loading. Duringthis period of time, the storage is carried out at a temperature of +5to 25° C. As may be easily appreciated, the adsorption percentage(adsorption rate) may be easily deduced from the desorption percentage(or desorption rate).

When the device is used not only to close the container but also todeliver the composition contained in the container, such as by slidingthe plunger of a syringe, it is recommended to siliconize the innersurface of the container.

However, it has been observed that silicone may in some cases bedetrimental to adsorption. Indeed, the desorption rate observed incompositions stored in syringes conventionally siliconized by meresurface-treatment with a silicone-in-water emulsion may be higher thanthe desorption rate observed in compositions stored in non-siliconizedcontainers. We postulate that although the silicone adheres to the innersurface of the container, it remains in free form and, upon shaking orstirring, can flow away from the inner surface and pass into thecontainer's content (the vaccine composition).

We have now found that this latter problem can be solved by using acontainer wherein the inner surfaces are coated with polymerizedsilicone. Such a container can be obtained by treating the inner surfaceof the container with a silicone-in-water emulsion, followed by heatingthe container, for example at a temperature of 270 to 330° C. for 30min. Upon heating, the silicone polymerizes on the inner surface of thecontainer and is therefore no longer capable of mixing with thecomposition. Polymerizing the silicone makes it possible to reduce thesurface energy of the silicone to which the vaccine composition may besensitive.

Additionally, the siliconizing operation comprising a polymerizationstep (i) is more precise and more homogeneous that a simple standardsiliconizing operation; and (ii) makes it possible to reduce the amountof silicone that is used (that is, loaded on the inner surface of thecontainer) by about a factor of 10 without any loss of lubricatingeffect. For example, according to a standard siliconizing process, from400 to 1000 μg of silicone are deposited in a syringe intended tocontain doses of 0.5-1 ml (the total inner surface of the 0.5-1 mlsyringe reservoir is about 8 cm²; in this example this surfacecorresponds to an amount of silicone of from about 50 to 125 μg/cm²),whereas from 40 to 100 μg of silicone are sufficient for the samesyringe (about 5 to 12 μg/cm²) if silicone is deposited on the innersurfaces of the container and then polymerized, for example by heating.The fact that the inner surface of the syringe is coated with a lowamount of polymerized silicone in a more homogenous manner than with alow amount of free silicone allows non-siliconized plungers to slidesmoothly, whereas such plungers are inoperative with syringes coatedwith low amount of free silicone.

This is the reason why the invention also comprises a second method forreducing and/or slowing down the desorption of an antigen adsorbed on analuminum adjuvant, the method comprising filling a container with avaccine composition comprising the antigen adsorbed on the aluminumadjuvant, wherein the inner surface of the container is coated withpolymerized silicone.

In other words, the invention also comprises a second method for loadingand storing a vaccine composition containing an antigen adsorbed on thealuminum adjuvant, the method comprising filling a container with thecomposition wherein the inner surface of the container is coated withpolymerized silicone.

In a similar manner, the invention also comprises the use of a containerhaving an inner surface coated with polymerized silicone for storing avaccine composition comprising an antigen adsorbed on an aluminumadjuvant.

Advantageously, the container having inner surfaces coated withpolymerized silicone is made of plastic or glass. Advantageously, thecontainer is the reservoir of a syringe.

The amount of polymerized silicone coated on the inner surface of thecontainer is from 3 to 25 μg/cm²; advantageously from 5 to 20 μg/cm²;preferably from 5 to 15 μg/cm².

As may be easily understood, the container used in the first methods ofthe invention may be advantageously coated with polymerized silicone asdescribed above.

In another aspect, the invention comprises:

A—A container (i) which contains a vaccine composition comprising anantigen adsorbed on an aluminum adjuvant; and (ii) which is closed by adevice acting as a stopper, wherein the surface of the devicecontactable with the composition is coated with a fluoropolymer;

B—A container (i) having inner surfaces coated with polymerizedsilicone; and (ii) containing a vaccine composition comprising anantigen adsorbed on an aluminum adjuvant; and

C—A container (i) having inner surfaces coated with polymerizedsilicone; (ii) containing a vaccine composition comprising an antigenadsorbed on an aluminum adjuvant; and (iii) which is closed by a deviceacting as a stopper, wherein the surface of the device contactable withthe composition is coated with a fluoropolymer.

In other words, the invention relates to a vaccine compositioncomprising an antigen adsorbed on an aluminum adjuvant which is loadedinto and stored in a container (i) having the inner surface coated withpolymerized silicone; and/or (ii) which is closed by a device acting asa stopper, wherein at least the surface of the device contactable withthe composition is coated with a fluoropolymer.

Vaccine compositions stored in containers according to the inventioninclude:

-   -   A vaccine composition comprising an antigen, wherein the minimal        antigen amount required for intended use (e.g., as a dose for        administration to a human) is adsorbed on an aluminum adjuvant;    -   A vaccine composition comprising an antigen adsorbed on an        aluminum adjuvant, wherein the vaccine composition when loaded        in a container for use in the present invention, exhibits an        adsorption percentage of at least:        -   (a) 65-70% of the total antigen amount present in the            composition, when, immediately after loading, the            composition is stored at (i) 5±3° C. for 2-3 years; or (ii)            25±3° C. for 2-3 months; or        -   (b) 80-90%, of the total antigen amount present in the            composition, when, immediately after loading, the            composition is stored at 5±3° C. for 18 months; and    -   A vaccine composition comprising an aluminum adjuvant and an        antigen able to adsorb onto the aluminum adjuvant, wherein the        percentage of the antigen adsorbed on the aluminum adjuvant is        at least 5 or 10% higher than the percentage observed when the        same vaccine composition is contained in a standard container        having uncoated inner surfaces or inner surfaces coated with        non-polymerized silicone, the comparison between the antigen        adsorption percentages being carried out after storage of the        vaccine-containing containers at 25° C. for 2 months, starting        from the date of filling the containers.

For the purposes of the present invention, the container may be any typeof reservoir, such as vials or syringes, and may contain multiple doses(multidose container) or a single dose (single-dose container). As anexample, the container may be a syringe or a part of a syringecomprising the reservoir containing the vaccine closed by a deviceacting as a stopper and as a system for releasing the vaccine at thetime of administration (e.g., using a plunger). The device acting as astopper may be a plunger.

Stoppers and/or plungers for use in the present invention are sold, forexample, by Helvoet Pharma (Omniflex™ technology) and by West Pharma(Fluorotec™ technology). Glass syringe reservoirs coated withpolymerized silicone for use in the present invention are sold, forexample, by Nuova Ompi srl, Becton-Dickinson and Gerresheimer (Baked-ontechnology).

The antigen and the aluminum adjuvant may be any antigen and anyaluminum adjuvant provided, of course, that they are both capable ofinteracting with each other. Furthermore, it may easily be understoodthat the present invention is of particular interest for anantigen-aluminum adjuvant pair having a relatively weak interactionforce; the interaction force possibly depending on the environment. Thisinteraction force can be assessed according to a variety of tests. Forexample, an aluminum adjuvant may be used to form variousantigen-adjuvant pairs (the antigen varies from one pair to the other,the adjuvant remaining the same). Then a large amount of a compound ableto compete with the antigen for the interaction with the adjuvant isadded. The various preparations are centrifuged and the supernatantsrecovered. Finally, the amount of antigen desorbed is assayed in thesupernatants, and as a result, antigens may be compared for theirinteraction force with the adjuvant.

A relatively weak interaction force is an interaction force that leadsto an adsorption that may be detrimentally affected by a standardfilling with the composition containing the antigen-aluminum adjuvantcomplex. Various elements involved in the manufacture of a container,such as, for example, latex, antioxidants, silicone and metal ions(e.g., zinc and tungsten), can destabilize the antigen-adjuvant complex.

By “adsorption” it is generally meant any phenomenon aimed at forming anantigen-adjuvant complex involving i.a. electrostatic interactionforces, hydrophobic interactions or ligand exchange. Thus, the antigenmay be attached at the surface of the network of the aluminum adjuvantor embedded inside after co-precipitation with the aluminum adjuvant.

For use in the present invention, an aluminum adjuvant may be aluminumoxy hydroxide (AlOOH), such as the product sold by Brenntag AG(Superfos) or Reheis Corp.; and aluminum hydroxy phosphate (AlOHPO₄),such as the product sold by Alphos.

For a vaccine composition to be effective, the minimal antigen amountrequired for adsorption onto the aluminum adjuvant essentially dependsupon the antigen itself, and is readily determinable by those ofordinary skill in the art.

In a particular embodiment, the antigen can be the hepatitis B surfaceantigen (HBsAg). It is particularly advantageous to adsorb HBsAg ontoAlOOH as HBsAg exhibits an iso electric point (IEP) less than 7 (about 4to 5) and AlOOH exhibits a point of zero charge (PZC) greater than 7(about 9 to 11).

The vaccine composition for use in the present invention can contain oneor more antigen(s), at least one of them being adsorbed on the aluminumadjuvant and it being possible for the others to be adsorbed as well ornot.

According to one embodiment, the composition for use in the presentinvention comprises HBsAg adsorbed on AlOOH (AlOOH-HbsAg complex) and asecond antigen, which is polyribosylribitol phosphate (PRP) ofHaemophilus influenzae (HiB valence), preferably in a form conjugated toa carrier protein (C) which may be i.a. Dt or Tt.

According to another particular embodiment, when the antigen adsorbed onthe aluminum adjuvant is HBsAg, the vaccine composition may alsocontain, as additional antigens, one or more of the following:diphtheria toxoid (Dt) (diphtheria valence); tetanus toxoid (Tt)(tetanus valence); Bordetella pertussis detoxified toxin (Ptdx),fimbriae, filamentous haemagglutinin (FHA) and/or pertactin (69 kDantigen) (pertussis valence); inactivated poliovirus serotype 1, 2, or 3(polio valence); and polyribosylribitol phosphate (PRP) of Haemophilusinfluenzae (HiB valence), preferably in a form conjugated to a carrierprotein (C) which may be i.a. Dt or Tt.

As a matter of example, the composition may comprise HBsAg, Dt, Tt, Ptand FHA adsorbed on AlOOH (the AlOOH-HbsAg-Dt-Tt-Pt-FHA complex), thepolio valence, and PRP-C substantially non-adsorbed on AlOOH.

In a general manner, the invention is also particularly advantageouswhen the vaccine composition comprises several valences, for example 2,3, 4, 5, 6 or more, each represented by one or more antigens (2, 3, 4 or5), several antigens being adsorbed on the aluminum adjuvant. Indeed,under the standard filling mode, the higher the number ofantigens/valences adsorbed on the aluminum adjuvant, the more criticalis the phenomenon of destabilization by the container. Theantigen-adjuvant interaction force often differs from one antigen toanother and, in a composition containing several antigens, the antigenwith the weakest interaction force exhibits the highest tendency todesorb under adverse conditions.

Examples and Experimental Results

A—A bulk of a vaccine composition containing the hepatitis B surfaceantigen, diphtheria toxoid, tetanus toxoid, and pertussis valence, eachadsorbed on aluminum oxy hydroxyde, as well as the polio andnon-adsorbed Haemophilus influenzae B (HiB) valences was distributedinto three categories of single-dose syringes, the characteristics ofwhich were as follows:

-   -   (1) standard siliconized glass syringes with standard        stopper/plunger made of non-laminated plastic;    -   (2) standard siliconized glass syringes with        fluoropolymer-coated stopper/plunger (West Pharma; Fluorotech™        technology); and    -   (3) syringes, the inner surface of which is coated with        polymerized silicone (Baked-on syringe system Luercone™ from        Gerresheimer) with fluoropolymer-coated stopper/plunger (West        Pharma; Fluorotech™ technology).

The bulk was distributed in 0.5 mL single doses, each dose containing 10μg of HBsAg, 30 Lf of Dt, 10 Lf of Tt, 25 μg of Pt, 25 μg of FHA, 40 DU(Antigen D Unit) of IPV1, 8 DU of IPV2, 32 DU of IPV3, 12 μg of PRP (inPRP-Tt conjugate form), 0.6 mg of Al, 55 mM of phosphate ions, 20 mM ofcarbonate ions, and Tris sucrose buffer, 2.5 mM, 2.125%, at pH 6.8-7.2.

All the syringes of the three categories were stored horizontally at 25°C. for two months (accelerated ageing). The HBsAg desorption wasmeasured in each of the three categories at T=0 (just after loading thesyringes) and then after two months.

Desorption was evaluated by centrifuging the content of the syringes andthen measuring the amount of desorbed HBsAg present in the supernatantby ELISA (sandwich ELISA, involving a mouse anti-HBsAg monoclonalantibody (IgM) for coating and capture, a second mouse anti-HBsAgmonoclonal antibody (IgG) and a third anti-mouse IgG polyclonal antibodycoupled to peroxydase (Sigma, Ref. A3673) which is revealed by addingtetramethyl benzidine).

At T (time)=0, the HbsAg adsorption level was identical in the threecategories (98% of the total HBsAg was adsorbed). At T=2 months,desorption was observed in all the categories, but the desorptionpercentage differed depending upon the category. The highest desorptionpercentage was found in category (1) (At T=1 and 2 months, 55 and 50% ofthe total HBsAg was adsorbed, respectively), whereas the lowestpercentage was found in category (3) (At T=1 and 2 months, 72 and 69% ofthe total HBsAg was adsorbed, respectively).

B—A bulk of the vaccine composition described in A—was distributed intotwo categories of single-dose 1 mL syringes, the characteristics ofwhich were as follows:

-   -   (1) standard siliconized glass syringes (free silicone); and    -   (2) non-siliconized syringes.

The bulk was distributed in 0.5 mL single doses, each dose containing 10μg of HBsAg, 30 Lf of Dt, 10 Lf of Tt, 25 μg of Pt, 25 μg of FHA, 40 DU(Antigen D Unit) of IPV1, 8 DU of IPV2, 32 DU of IPV3, 12 μg of PRP (inPRP-Tt conjugate form), 0.6 mg of Al, 55 mM of phosphate ions, 20 mM ofcarbonate ions, and Tris sucrose buffer, 2.5 mM, 2.125%, at pH 6.8-7.2.

All the syringes of the two categories were stored vertically at 25° C.for two months (accelerated ageing). The HBsAg desorption was measuredin each of the two categories at T=0 (just after filling syringes) andthen after two months, as described in A—above.

At T (time)=0, the HbsAg adsorption level was identical in the twocategories (98% of the total HBsAg was adsorbed). At T=2 months,desorption was observed in all the categories, but the desorptionpercentage differed depending upon the category. The highest desorptionpercentage was found in category (1) (At T=1 and 2 months, 69 and 68% ofthe total HBsAg was adsorbed, respectively), whereas the lowestdesorption percentage was found in category (2) (At T=1 and 2 months,73% of the total HBsAg was adsorbed). This clearly indicates that theantigen adsorption onto an aluminum adjuvant is sensitive to freesilicone.

C—A bulk of the vaccine composition described in A—was distributed intothree types of single-dose 1 mL syringes, the characteristics of whichwere as follows:

-   -   (1) low siliconized glass syringes (50-100 μg free        silicone/syringe);    -   (2) standard highly siliconized glass syringes ((RTF syringe        Luercone™ from Gerresheimer): 800 μg to 1 mg free        silicone/syringe); and    -   (3) syringes having inner surface coated with polymerized        silicone (50-100 μg/syringe).

Syringes of category (1) are operative only if the plunger used forinjection is also siliconized, because the amount of silicone coatingthe inner surface of the syringe is too low to allow sliding on its own.

All the syringes (types 1 to 3) were closed with the same type ofnon-siliconized stopper.

The bulk vaccine was distributed in 0.5 mL single doses, each dose beingas described in A and B.

All the syringes of the three types were stored vertically at 25° C. fortwo months (accelerated ageing). The HBsAg desorption was measured ineach of the three types at T=0 (just after filling syringes) and thenafter two months, as described in A—above.

At T (time)=0, the HbsAg adsorption level was identical in the threetypes (94% of the total HBsAg was adsorbed). At T=2 months, desorptionwas observed in all syringes, but the desorption percentage differeddepending upon the type. The highest desorption percentage was found intype (2) (At T=1 and 2 months, 60 and 58% of the total HBsAg wasadsorbed, respectively), whereas the desorption percentage was foundsimilar in types (1) and (3) and definitively much lower than in type(2): In type (1), at T=1 and 2 months, 73% and 68% of the total HBsAgwas adsorbed, respectively. In type (3), at T=1 and 2 months, 69% and66% of the total HBsAg was adsorbed, respectively.

Again, this clearly indicates that (i) the antigen adsorption onto analuminum adjuvant is sensitive to free silicone loaded in an amountnecessary for sliding and (ii) polymerization of silicone allowsovercoming this issue.

D—A stability study has been conducted at 5±3° C. for 18 months with thefilled syringes described in A—(3). At least 80-90% of the total HBsAgwas still adsorbed at the end of the 18-month period.

What is claimed is:
 1. A vaccine comprising a container containing aliquid multivalent vaccine composition comprising hepatitis B surfaceantigen adsorbed on aluminum oxyhydroxide, wherein the container (i) hasinner surfaces coated with polymerized silicone; and (ii) is closed by adevice acting as a stopper, wherein the surface of the devicecontactable with the vaccine composition is coated with a fluoropolymer,wherein the vaccine composition has reduced desorption rate of hepatitisB as compared to an otherwise identical vaccine composition stored in astandard container not having inner surfaces being coated withpolymerized silicone and/or closed by a device coated with the saidfluoropolymer.
 2. The vaccine as claimed in claim 1, wherein thefluoropolymer is selected from the group consisting ofpolytetrafluoroethylene (PTFE), polytetrafluoropropylene (PTFP),fluorinated ethylene propylene (FEP, a copolymer of hexafluoropropyleneand tetrafluoroethylene), polychlorotrifluoroethylene (PCTFE),perfluoroalkoxy co-polymer (PFA), poly(ethylene-co-tetrafluoroethylene)(ETFE), poly(ethylenechlorotrifluoroethylene) (ECTFE), polyvinylfluoride (PVF) and polyvinylidene fluoride (PVPF).
 3. The vaccine asclaimed in claim 1, wherein the container is a syringe and the deviceacting as a stopper is a plunger.
 4. The vaccine as claimed in claim 1,wherein the percentage of the hepatitis B adsorbed on the aluminumadjuvant is at least 5% higher than that of the otherwise identicalvaccine composition stored in the standard container, when storage iscarried out at 25° C. for 2 months from date of filling.
 5. The vaccineas claimed in claim 1, wherein the desorption rate is reduced by 10% ormore as compared to an otherwise identical vaccine composition stored inthe standard container, when storage is carried out at 5-25° C. for 1 or2 months from date of filling.
 6. The vaccine as claimed in claim 1,wherein the inner surfaces of the container are coated with polymerizedsilicone at an amount from 3 to 25 μg/cm².
 7. The vaccine as claimed inclaim 1, wherein the vaccine composition further comprises additionalantigens selected from one or more of diphtheria toxoid (Dt) (diphtheriavalence); tetanus toxoid (Tt) (tetanus valence); Bordetella pertussisdetoxified toxin (Ptdx), fimbriae, filamentous haemagglutinin (FHA)and/or pertactin (69 kD antigen) (pertussis valence); inactivatedpoliovirus serotype 1, 2, or 3 (polio valence); or polyribosylribitolphosphate (PRP) of Haemophilus influenzae (HiB valence).
 8. The vaccineas claimed in claim 1, wherein the vaccine is a single-dose vaccine. 9.The vaccine as claimed in claim 8, wherein each single-dose comprises 10μg of hepatitis B surface antigen.
 10. The vaccine as claimed in claim7, wherein each single-dose comprises 0.6 mg aluminum adjuvant.
 11. Thevaccine as claimed in claim 2, wherein the fluoropolymer ispoly(ethylene-co-tetrafluoroethylene) (ETFE).
 12. The vaccine as claimedin claim 2, wherein the fluoropolymer is polytetrafluoroethylene (PTFE).13. A vaccine comprising a container containing a liquid vaccinecomposition comprising hepatitis B surface antigen adsorbed on analuminum adjuvant, wherein the container (i) has inner surfaces coatedwith polymerized silicone at an amount from 3 to 25 μg/cm²; and (ii) isclosed by a device acting as a stopper, wherein the surface of thedevice contactable with the vaccine composition is coated with afluoropolymer, wherein the vaccine composition has reduced desorptionrate of hepatitis B as compared to an otherwise identical vaccinecomposition stored in a standard container not having inner surfacesbeing coated with polymerized silicone and/or closed by a device coatedwith the said fluoropolymer.